The ciliopathies are an emerging group of clinically overlapping disorders, hallmarked by features that include retinal degeneration, renal cystic disease, and central and peripheral nervous system defects. The unification of diverse clinical phenotypes such as nephronophthisis, Bardet-Biedl, Meckel-Gruber and Jeune syndromes is driven by both common cellular basis (defects at the primary cilium) and genetic overlap, where mutations in the same genes can both cause discrete phenotypes and contribute modifying mutations that modulate the penetrance and expressivity of primary genetic lesions. These observations have led to the idea that the total mutational load in the primary cilium is a strong phenotypic determinant. Moreover, robust, physiologically relevant in vitro and in vivo assays that report on organelle output affords us the opportunity to study the nature and properties of second-site modifiers in humans. During the first funding period, we established the ciliopathies as a unified entity, identified several causal and modifying genes and alleles, and developed assays to evaluate their effect. We now propose to expand on this work and, for the first time, assess the total mutational load in a discrete, biochemically characterized macromolecular system ciliary functional system. We will 1) sequence a large cohort of patients across the severity spectrum for all known intraflagellar transport genes and ask how phenotypic severity might track with genetic lesions;2) we will functionally assay all discovered alleles using a combination of in vivo complementation and in vitro protein stability and localization assays in ciliated cells;3) finally, we will begin to interrogate the significance of such alleles in mammals. As part of our Preliminary Data, we have identified a missense allele, P209L, in IFT139 that appears genetically sufficient to cause cystic renal disease but also interacts with alleles in other ciliopathy genes as a potential cystogenic modifier. We will therefore engineer a knock-in mouse and ask a) whether homozygosity for this mutation is sufficient for renal cyst formation;and b) whether introduction of 209L in a genetically sensitized background can exacerbate or potentiate cystic renal disease. Our studies will yield additional loci that cause ciliary disease and expand our knowledge base of second-site modifiers. Moreover, we anticipate that understanding of the genetic attributes of epistasis has the potential to inform a broad range of disorders and improve the clinical utility of genetic information.
Although individually rare, the ciliopathies collectively represent a significant health burden and their genetic dissection will expedite prognosis, management, and treatment. Moreover, this group of disorders is emerging as an exceptionally useful system to study the relationship between genetic load in a functional system and clinical variability, offering a unique opportunity to understand pathology and disease progression at the level of the individual patient.
|Katsanis, Nicholas (2018) Point: Treating Human Genetic Disease One Base Pair at a Time: The Benefits of Gene Editing. Clin Chem 64:486-488|
|Heydeck, Westley; Fievet, Lorraine; Davis, Erica E et al. (2018) The complexity of the cilium: spatiotemporal diversity of an ancient organelle. Curr Opin Cell Biol 55:139-149|
|Liu, Yangfan P; Bosch, Daniëlle G M; Siemiatkowska, Anna M et al. (2017) Putative digenic inheritance of heterozygous RP1L1 and C2orf71 null mutations in syndromic retinal dystrophy. Ophthalmic Genet 38:127-132|
|Goetz, Sarah C; Bangs, Fiona; Barrington, Chloe L et al. (2017) The Meckel syndrome- associated protein MKS1 functionally interacts with components of the BBSome and IFT complexes to mediate ciliary trafficking and hedgehog signaling. PLoS One 12:e0173399|
|Helm, Benjamin M; Willer, Jason R; Sadeghpour, Azita et al. (2017) Partial uniparental isodisomy of chromosome 16 unmasks a deleterious biallelic mutation in IFT140 that causes Mainzer-Saldino syndrome. Hum Genomics 11:16|
|Frosk, Patrick; Arts, Heleen H; Philippe, Julien et al. (2017) A truncating mutation in CEP55 is the likely cause of MARCH, a novel syndrome affecting neuronal mitosis. J Med Genet 54:490-501|
|Prieto-Echagüe, Victoria; Lodh, Sukanya; Colman, Laura et al. (2017) BBS4 regulates the expression and secretion of FSTL1, a protein that participates in ciliogenesis and the differentiation of 3T3-L1. Sci Rep 7:9765|
|Ta-Shma, Asaf; Khan, Tahir N; Vivante, Asaf et al. (2017) Mutations in TMEM260 Cause a Pediatric Neurodevelopmental, Cardiac, and Renal Syndrome. Am J Hum Genet 100:666-675|
|Dougherty, Gerard W; Loges, Niki T; Klinkenbusch, Judith A et al. (2016) DNAH11 Localization in the Proximal Region of Respiratory Cilia Defines Distinct Outer Dynein Arm Complexes. Am J Respir Cell Mol Biol 55:213-24|
|Boldt, Karsten; van Reeuwijk, Jeroen; Lu, Qianhao et al. (2016) An organelle-specific protein landscape identifies novel diseases and molecular mechanisms. Nat Commun 7:11491|
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